Comprehensive detection of viruses in pediatric patients with acute liver failure using next-generation sequencing |
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| Affiliation: | 1. Departments of Pediatrics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 466-8550, Japan;2. Department of Pediatric Hepatology and Gastroenterology, Saiseikai Yokohamashi Tobu Hospital, 3-6-1 Shimosueyoshi, Tsurumi-ku, Yokohama, Kanagawa 230-0012, Japan;1. Research and Development Institute, National Infection Service, Public Health England, Porton Down, Salisbury, Wilts SP40JG, UK;1. Veterinary and Agrochemical Research Centre CODA/CERVA, Groeselenberg 99, Brussels Belgium;2. The Pirbright Institute, Ash Rd, Woking GU24 0NF, United Kingdom;1. Karadeniz Technical University, Faculty of Sciences, Department of Molecular Biology and Genetic, 61080 Trabzon, Turkey;2. Karadeniz Technical University, Faculty of Sciences, Department of Biology, 61080 Trabzon, Turkey;3. Laboratory for Molecular Virology, Great Lakes Forestry Centre, Sault Ste. Marie, Ontario, Canada;1. Department of Pediatrics, Nagoya University Graduate School of Medicine, Nagoya, Japan;2. Department of Genetics, Research Institute of Environmental Medicine, Nagoya University, Nagoya, Japan;3. Division of Clinical Microbiology, Department of Laboratory Medicine, Karolinska Institute, Stockholm, Sweden;4. Department of Pediatrics and Child Health, Nihon University School of Medicine, Tokyo, Japan;1. Department of Marine Life Sciences, Jeju National University, Jeju Self-Governing Province 63243, Republic of Korea;2. Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province 63243, Republic of Korea |
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| Abstract: | BackgroundPediatric acute liver failure (PALF) is a rare and severe syndrome that frequently requires liver transplantation. Viruses are one of the most frequent causes of this disease, however, pathogenic viruses are not determined in many patients. Recently next-generation sequencing (NGS) has been applied to comprehensively detect pathogens of infectious diseases of unknown etiology.ObjectivesTo evaluate an NGS-based approach for detecting pathogenic viruses in patients with PALF or acute hepatitis of unknown etiology.Study designTo detect virus-derived DNA and RNA sequences existing in sera/plasma from patients, both DNA and RNA sequencing were performed. First, we validated the ability of NGS to detect viral pathogens in clinical serum/plasma samples, and compared different commercial RNA library preparation methods Then, serum/plasma of fourteen patients with PALF or acute hepatitis of unknown etiology were evaluated using NGS.ResultsAmong three RNA library preparation methods, Ovation RNA-Seq System V2 had the highest sensitivity to detect RNA viral sequences. Among fourteen patients, sequence reads of torque teno virus, adeno-associated virus, and stealth virus were found in the sera of one patient each, however, the pathophysiological role of these three viruses was not clarified. Significant virus reads were not detected in the remaining 11 patients.ConclusionsThis finding might be due to low virus titer in blood at the time of referral or a non-infectious cause might be more frequent. These results suggest an NGS-based approach has potential to detect viral pathogens in clinical samples and would contribute to clarification of the etiology of PALF. |
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| Keywords: | Next-generation sequencing Acute liver failure Acute hepatitis Pathogen |
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